Showing papers by "Nagoya Institute of Technology published in 2008"
03 Sep 2008
TL;DR: This work presents a method for predicting diffusion probabilities by using the EM algorithm, and defines the likelihood for information diffusion episodes, where an episode means a sequence of newly active nodes.
Abstract: We address a problem of predicting diffusion probabilities in complex networks. As one approach to this problem, we focus on the independent cascade (IC) model, and define the likelihood for information diffusion episodes, where an episode means a sequence of newly active nodes. Then, we present a method for predicting diffusion probabilities by using the EM algorithm. Our experiments using a real network data set show the proposed method works well.
463 citations
TL;DR: In this article, the authors used discrete element models to show the dependence of critical state conditions on grading and the way in which the particle assembly seeks out new critical state condition as the grading changes.
Abstract: Examples of situations are presented where the grading of a soil changes during its lifetime either by crushing of particles leading to an increase of fine material or by slow transport of fine particles with seepage leading to a decrease of fine material. Such grading changes influence the basic constitutive properties of the soil, in particular properties such as critical states which are dependent on the available range of densities of packing. Discrete element modelling is used to show the dependence of critical state conditions on grading and the way in which the particle assembly seeks out new critical state conditions as the grading changes.
345 citations
289 citations
TL;DR: In this paper, the authors discuss recent developments in the enantioselective trifluoromethylation reactions, including nucleophilic, electrophilic, and radical approaches.
Abstract: Fluorinated organic compounds have received considerable attention because they possess unique properties such as greater stability, higher electronegativity, and water and oil repellency, which are potentially beneficial in terms of the development of novel pharmaceutical agents and advanced materials. Such extreme behaviors of fluoro-organic compounds however, often make their syntheses all the more complicated, especially when the synthesis of target compounds requires an enantiocontrolled methodology. Many strategies can be employed for introducing chirality into organic molecules, but enantioselective fluorination and trifluoromethylation reactions are especially attractive because non-fluorinated prochiral substrates can be directly transformed to chiral fluoro-organic compounds with a control of absolute configuration. Significant progress has been made in the development of the asymmetric fluorination reaction in recent years; however, the direct enantioselective trifluoromethylation reaction remains a challenge. In this report, we discuss recent developments in the enantioselective trifluoromethylation reactions, including nucleophilic, electrophilic, and radical approaches. Although the emphasis of this report is on the enantioselective trifluoromethylation reaction, the less-studied enantioselective difluoromethylation and monofluoromethylation reactions are also covered.
275 citations
TL;DR: Experimental results demonstrate that the MLE- based mapping with dynamic features can significantly improve the mapping performance compared with the MMSE-based mapping in both the articulatory-to-acoustic mapping and the inversion mapping.
251 citations
TL;DR: In this article, gold-coated silver and silvercoated gold composite nanoparticles are prepared by a seeding growth method to study their SERS-active properties, which integrates high levels of sensitivity with spectroscopic precision and has tremendous potential for chemical and biomolecular sensing.
228 citations
TL;DR: Co-doped silica sol solutions with varying Co composition (Co/(Si+Co)=10−50 mol%) were prepared from tetraethoxysilane and Co(NO3)2·6H2O as discussed by the authors.
Abstract: Co-doped silica sol solutions with varying Co composition (Co/(Si+Co)=10–50 mol%) were prepared from tetraethoxysilane and Co(NO3)2·6H2O. Subsequently, these solutions were used in the preparation of hydrogen separation microporous membranes with enhanced hydrothermal stability at 500°C under a steam pressure of 300 kPa. At Co concentrations >33%, the XRD pattern and peak intensity of the Co-doped silica preparations were similar and were not dependent on Co composition, suggesting that Co was incorporated into the silica network. The best H2 permeation performance in a steam atmosphere (500°C; steam pressure, 300 kPa) was obtained using silica doped with approximately 30 mol% Co. Co-doped silica membranes (Co 33 mol%) fired at 600°C under a steam partial pressure of 90 kPa showed stable gaseous permeances and a H2 permeance of approximately 2.00–4.00 × 10−6 m3(STP)·(m·s·kPa)−1 with a selectivity of 250–730 (H2/N2), even after 60 h of exposure to steam (steam pressure, 300 kPa) at 500°C.
169 citations
164 citations
TL;DR: In this article, a Johnson-type methyl transfer reagent was designed for electrophilic trifluoromethylation of carbon nucleophiles, which was used for the first time for dicyanoalkylidenes.
151 citations
TL;DR: In this article, microstructural evolution taking place during equal channel angular pressing (ECAP) was studied in a coarse-grained Al-6% Mg-0.4% Mn-Sc alloy at 300°C (∼0.6 T m ).
139 citations
TL;DR: Novel natural N-heteroarylsulfonylprolinamide organocatalysts are designed, which would enable the TS to be controlled by intramolecular hydrogen bonding between the sulfonimide NH proton and the heteroatom of the heteroaryl group.
Abstract: Convolutamydine A is an alkaloid that was isolated from the Floridian marine bryozoan Amathia convoluta by Kamano and co-workers in 1995. (R)-Convolutamydine A exhibits a potent inhibitory activity towards the differentiation of HL-60 human promyelocytic leukaemia cells at 0.1– 25 mgmL . In 2006, Gargen and Tomasini0s group reported the first enantioselective synthesis of natural (R)-convolutACHTUNGTRENNUNGamydine A through a direct aldol reaction of 4,6-dibromoACHTUNGTRENNUNGisatin with acetone by using 10 mol% of a peptidic catalyst derived from d-proline and l-b-homophenylgricine. Very recently, Xiao and co-workers reported the same reaction with 20 mol% of a chiral bisamide, prepared from a chiral diamine and l-proline, as an organocatalyst to give 60% enantionmeric excess (ee) of the unnatural (S)-convolutamydine A. Malkov and co-workers reported the synthesis of (R)-convolutamydine A with high enantiopurity (up to 94% ee) by using 20 mol% of d-leucinol prepared from unnatural d-leucine; however, it takes a long time (RT, 36 h) for the reaction to reach completion. Unsolved problems include high catalyst loading and the use of organocatalysts derived from unnatural a-amino acids to prepare (R)-convolutamydine A. On the other hand, organocatalytic aldol reactions with ketones as acceptors generally need a high catalyst loading (at least 5–10 mol%) of organocatalyst, whereas enantioselective aldol reactions with aldehydes have been achieved with low catalyst loading of organocatalysts. Recently, we and others have reported enantioselective reactions that use 2-point coordinative heteroarylsulfonyl groups, which can control the transition state (TS) or intermediates by chelation with chiral Lewis acids or organocatalysts. To overcome the difficulty of the enantioselective reaction between 4,6-dibromoisatin as the ketone acceptor and acetone, we designed novel natural N-heteroarylsulfonylprolinamide organocatalysts, which would enable the TS to be controlled by intramolecular hydrogen bonding between the sulfonimide NH proton and the heteroatom of the heteroaryl group (Scheme 1). Herein, we report the highly efficient synthesis of (R)-convolutamydine A and its derivatives by using a catalytic amount of novel N-heteroarylsulfonylprolinamide organocatalyst.
TL;DR: The first catalytic, asymmetric 1,4-conjugate addition of FBSM to a,b-unsaturated ketones is reported, which is one of the most powerful tools for carbon–carbon bond-forming reactions.
Abstract: There is a high demand in both academia and industry for enantiopure fluorine-containing organic molecules because of their unique pharmacological properties. Although their occurrence in natural systems is rare, monofluorinated analogues of biologically active compounds are often evaluated as bioisosteres of the parent molecules. Compounds with monofluoromethyl groups are also important in biological systems. The current state-of-the-art asymmetric catalysis uses organocatalysts or ligand–metal complexes that allow access to the chiral monofluorinated organic compounds with high enantiocontrol. However, most of the recent innovations in this area are based on the enantioselective fluorination reactions developed by us and others. In comparison, the enantioselective fluoromethylation reactions still need to be investigated. In 2006, we disclosed 1fluorobis(phenylsulfonyl)methane (FBSM) as a synthetic equivalent of a monofluoromethide species under the palladium-catalyzed Tsuji–Trost allylic alkylation conditions, which provided the first asymmetric allylic monofluoromethylation with high enantiocontrol. Hu and co-workers, and Prakash, Olah, and co-workers demonstrated the effectiveness of FBSM for achieving monofluoromethylation in the non-asymmetric epoxide ring-opening and Mitsunobu reactions. Recently, we reported the FBSM-based catalytic enantioselective Mannich-type monofluoromethylation which provided chiral a-fluoromethylamines with excellent enantioselectivities. On the basis of this concept we used FBSM as a potential monofluoromethide equivalent, and report herein the first catalytic, asymmetric 1,4-conjugate addition of FBSM to a,b-unsaturated ketones. The ammonium salts of cinchona alkaloids possessing sterically demanding substituents effectively catalyzed the conjugate addition reaction to furnish Michael adducts in high yield with excellent enantioselectivity. The catalytic asymmetric Michael addition reaction is one of the most powerful tools for carbon–carbon bond-forming reactions. Either chiral quaternary ammonium salts or chiral Lewis acids can be successfully used as catalysts for achieving high enantiocontrol. There are many reports of the catalytic asymmetric conjugate addition of nucleophiles to a Michael acceptor; however, it has not been extended to asymmetric fluoromethylations. On the basis of our work on asymmetric monofluoromethylations, FBSM was envisioned to function as a Michael-type donor under suitable conditions. This assumption was first tested on the reaction of FBSM with chalcone (1a) under our best conditions for asymmetric Mannich-type fluoromethylation, which employed benzylquinidinium chloride (QD-3a ; X=Cl) as a catalyst (10 mol%) in the presence excess CsOH·H2O (3.0 equiv) in CH2Cl2 at low temperature. The initial results were quite discouraging as the reaction produced Michael adduct 2a in a 62% yield with a low ee value (Table 1, entry 1). The ee value was improved to 44% when the reaction was performed in the presence of K2CO3 (Table 1, entry 2). After screening various bases and solvents (Table 1, entries 3–8), a catalytic amount of QD-3a (X=Cl) and 3 equivalents of Cs2CO3 in CH2Cl2 provided (S)-2a with the highest enantioselectivity (Table 1, entry 4). Attempts to improve the enantioselectivity of the product by using catalyst QD-4 failed, providing 2a in a low yield with 27% ee (Table 1, entry 9). This finding indicated that the free hydroxy group of the cinchona alkaloid is indispensable for achieving high enantiocontrol. Ammonium salts, CN-3a and CD-3a, derived from cinchonine and cinchonidine, respectively, were less effective (Table 1, entries 11 and 12). An extensive screening of cinchona alkaloids was performed under the same conditions (Table 1, entries 10–17), and we found that the quinidinium chloride (QD-3 f ; X=Br), bearing a sterically demanding benzyl substituent, was effective at providing 2a in high yield with an excellent ee value of 97% (Table 1, entry 17). We also discovered that by using the analogous ammonium bromide derived from quinine (QN-3 f ; X=Br), a similar enantioselectivity was obtained for 2a, albeit with the opposite (R) stereochemistry (Table 1, entry 18). Notably, for the reaction of 1a with FBSM at low catalyst loading (5 mol%), the same high ee value of (S)-2a was obtained (Table 1, entry 19). [*] T. Furukawa, Prof. N. Shibata, Prof. S. Mizuta, Dr. S. Nakamura, Prof. T. Toru Department of Frontier Materials, Graduate School of Engineering Nagoya Institute of Technology Gokiso, Showa-ku, Nagoya, 466-8555 (Japan) Fax: (+81)52-735-5442 E-mail: nozshiba@nitech.ac.jp
TL;DR: It is shown that both genomic amplification and deletion strongly influence the expression of genes in altered genomic regions, and it is proposed that the list of candidate genes may contain novel genes involved in the pathogenesis of advanced gastric cancer.
Abstract: Genomic copy number aberrations (CNAs) are believed to play a major role in the development and progression of human cancers. Although many CNAs have been reported in gastric cancer, their genome-wide transcriptional consequences are poorly understood. In this study, to reveal the impact of CNAs on genome-wide expression in gastric cancer, we analysed 30 cases of gastric cancers for their CNAs by array comparative genomic hybridization (array CGH) and 24 of these 30 cases for their expression profiles by oligonucleotide-expression microarray. We found that with the application of laser microdissection, most CNAs were detected at higher frequency than in previous studies. Notably, gain at 20q13 was detected in almost all cases (97%), suggesting that this may play an important role in the pathogenesis of gastric cancer. By comparing the array CGH data with expression profiles of the same samples, we showed that both genomic amplification and deletion strongly influence the expression of genes in altered genomic regions. Furthermore, we identified 125 candidate genes, consisting of 114 up-regulated genes located in recurrent regions (>10%) of amplification and 11 down-regulated genes located in recurrent regions of deletion. Up-regulation of several candidate genes, such as CDC6, SEC61G, ANP32E, BYSL and FDFT1, was confirmed by immunohistochemistry. Interestingly, some candidate genes were localized at genomic loci adjacent to well-known genes such as EGFR, ERBB2 and SMAD4, and concordantly deregulated by genomic alterations. Based on these results, we propose that our list of candidate genes may contain novel genes involved in the pathogenesis of advanced gastric cancer.
TL;DR: Thermal decomposition behaviors of a series of imidazolium-type ionic liquid samples were studied by pyrolysis-gas chromatography at 550 degrees C using various detectors, finding that no boron-containing products were found even from the samples with BF(4) anion, whereas minor but clear peaks of phosphorous- containing products were observed in the pyrograms of the sample with PF(6) anions.
Abstract: Thermal decomposition behaviors of a series of imidazolium-type ionic liquid samples were studied by pyrolysis-gas chromatography at 550°C using various detectors. As for the imidazolium halides, haloalkanes and 1-alkylimidazoles corresponding to the alkyl substituents were mainly formed through the nucleophilic attacks of halide ions to the alkyl groups followed by C-N bond cleavage, along with a minor amount of alkenes. Meanwhile, in the case of the ionic liquids with BF4, PF6 and CF3SO3 anions, corresponding alkenes were predominantly produced along with 1-alkylimidazoles rather than haloalkanes. No boron-containing products were found even from the samples with BF4 anion, whereas minor but clear peaks of phosphorous-containing products were observed in the pyrograms of the samples with PF6 anions. As for the samples with longer alkyl group, the pyrolyzates reflecting the C-C bond scissions in the alkyl groups were also formed to some extent. Meanwhile, imidazole rings did not decompose under the experimental conditions at around 550°C.
TL;DR: In this paper, the magnetic and ferroelectric properties of multiferroic RMn2O5 (R = Y, Tb, Ho, Er, Tm) are reviewed based on recent neutron diffraction and dielectric measurements.
Abstract: The magnetic and ferroelectric properties of multiferroic RMn2O5 (R = Y, Tb, Ho, Er, Tm) are reviewed based on recent neutron diffraction and dielectric measurements. Successive phase transitions of magnetic and dielectric ordering were found to occur simultaneously in this system. The characteristic magnetic ordering of the system exhibits an incommensurate–commensurate phase transition, and again transitions to an incommensurate phase. Special attention is given to the magnetic structure in order to discuss the mechanism for the introduction of ferroelectric polarization. For all the compounds examined, the spin configuration for Mn4+ and Mn3+ ions in the commensurate magnetic phase, where spontaneous electric polarization occurs, was determined to be a transverse spiral spin structure propagating along the c-axis. By contrast, the alignment of the induced 4f moment of R3+ ions showed variation, depending on the character of each of the elements. Corresponding responses to external fields such as a magnetic field, hydrostatic pressure etc at low temperature are strongly dependent on the rare earth element present in the RMn2O5 system. The so-called colossal magnetoelectric effect in this system can be easily interpreted by the phase transition from the magnetic incommensurate and weak ferroelectric phase to the commensurate and ferroelectric phase.
TL;DR: Direct utilization of palm oil for the simultaneous production of polyhydroxyalkanoates (PHAs) and rhamnolipids was demonstrated using Pseudomonas aeruginosa IFO3924 using secreted lipase.
Abstract: Direct utilization of palm oil for the simultaneous production of polyhydroxyalkanoates (PHAs) and rhamnolipids was demonstrated using Pseudomonas aeruginosa IFO3924. By secreted lipase, palm oil was hydrolyzed into glycerol and fatty acids. Fatty acids became favorable carbon sources for cell growth and PHA production via β-oxidation and glycerol for rhamnolipid production via de novo fatty acid synthesis. Both PHA and rhamnolipid syntheses started after the nitrogen source was exhausted and cell growth ceased. PHA synthesis continued until all fatty acids were exhausted, and at that time, PHA content in the cells reached a maximum, but stopped despite the remaining glycerol (<2g/l). In contrast, rhamnolipid synthesis continued until glycerol was exhausted.
TL;DR: In this paper, a variety of indoles with ethyl 3,3,3-trifluoropyruvate catalyzed by novel chiral m-phenylene- bis(imidazoline)-copper(II) complexes or the bis-(imidrazoline)-achiral acid combination afforded products with high enantioselectivity.
Abstract: Enantioselective Friedel-Crafts alkylations of a variety of indoles with ethyl 3,3,3-trifluoropyruvate catalyzed by novel chiral m-phenylene- bis(imidazoline)-copper(II) complexes or the bis-(imidazoline)-achiral acid combination afforded products with high enantioselectivity. Both enantiomers of indole derivatives can be prepared with high enantioselectivities by tuning the N-substituents of the imidazoline.
TL;DR: Cucurbit[7]uril forms very strong complex with zwitterionic dipeptide Phe-Gly with affinity exceeding 10(7) M(-1) and effectively recognizes peptide sequence of Phe,Gly over Gly-Phe as well as Tyr, Tyr, Gly and Trp over Trp with relative affinities of 23 000, 18 000 and 2000, respectively.
TL;DR: In this article, the authors investigated bipolar resistance switching on sputtered Pr0.7Ca0.3MnO3 sandwiched by Pt-and Ti-electrodes and found that negative differential resistance observed in the forming process originates from the motion of oxygen ions at the Ti/PCMO interface.
Abstract: Bipolar resistance switching was investigated on sputtered Pr0.7Ca0.3MnO3 (PCMO) sandwiched by Pt- and Ti-electrodes. Based on electrical conductivity measurements and a combination of electron energy loss spectroscopy analysis and transmission electron microscopy observation, we found that the negative differential resistance observed in the forming process originates from the motion of oxygen ions at the Ti/PCMO interface. We propose that the observed resistance switching is caused by an oxidation/reduction reaction at the interface.
TL;DR: Rheological measurements reveal that the viscoelasticity of the material can be tuned by environmental pH and peptide concentration, and the self-assembled hydrogels are able to recover from mechanical breakdowns.
25 Jan 2008-Materials Science and Engineering A-structural Materials Properties Microstructure and Processing
TL;DR: Grain refinement taking place in a coarse grained 2219 aluminum alloy deformed by equal channel angular pressing (ECAP) at a temperature of 250°C to a strain of 12 was investigated in this article.
Abstract: Grain refinement taking place in a coarse grained 2219 aluminum alloy deformed by equal channel angular pressing (ECAP) at a temperature of 250 °C to a strain of 12 was investigated. The microstructural changes are mainly characterized by the evolution of high density dislocation substructures with relatively low misorientations (∼3–5°) at strains below 2 and deformation bands (DBs) with moderate misorientation angle, which are frequently formed at strains above 3. Repeated ECAP leads to increasing the number and misorientation angle of the boundaries of DBs, so that the misorientation distribution for newly evolved dislocation subboundaries changes from single peak type at low strains to bimodal one at high strains. The fraction of high angle boundaries starts to increase at strain ɛ = 3, then it rapidly rises to about 80% at moderate to high strains and a new fine grains with almost random crystal orientation are fully developed in a whole material. The main mechanism of new fine grained structure formation during ECAP is discussed.
TL;DR: The results indicate that both AFs and MTs play crucial roles in maintaining whole cell mechanical properties of SMCs, and that while AFs act as an internal tension generator, MTs acts as a tension reducer, and these contribute to intracellular force balance three dimensionally.
Abstract: The effects of actin filaments (AFs) and microtubules (MTs) on quasi-in situ tensile properties and intracellular force balance were studied in cultured rat aortic smooth muscle cells (SMCs). A SMC...
TL;DR: A statistical parametric speech synthesis system developed by a joint group from the Nagoya Institute of Technology and the Nara Institute of Science and Technology is described for the annual open evaluation of text-to-speech synthesis systems named Blizzard Challenge 2006.
Abstract: We describe a statistical parametric speech synthesis system developed by a joint group from the Nagoya Institute of Technology (Nitech) and the Nara Institute of Science and Technology (NAIST) for the annual open evaluation of text-to-speech synthesis systems named Blizzard Challenge 2006. To improve our 2005 system (Nitech-HTS 2005), we investigated new features such as mel-generalized cepstrum-based line spectral pairs (MGC-LSPs), maximum likelihood linear transform (MLLT), and a full covariance global variance (GV) probability density function (pdf). A combination of mel-cepstral coefficients, MLLT, and full covariance GV pdf scored highest in subjective listening tests, and the 2006 system performed significantly better than the 2005 system. The Blizzard Challenge 2006 evaluations show that Nitech-NAIST-HTS 2006 is competitive even when working with relatively large speech databases.
TL;DR: In this article, a broad photoluminescence band with a large Stokes shift is observed, and its decay time depends strongly on the emission energies, attributed to radiative recombination of photo-excited carriers after multiple-step hopping among deep trapping sites on nanoparticle surfaces.
TL;DR: In this article, X-ray diffraction, scanning electron microscopy and specific surface area measurements were used to analyze the structural and microstructural changes of the as prepared powders heated at different temperatures.
Abstract: Lanthanum silicate oxyapatites of structural formula La9.33+x(SiO4)6O2+1.5x (0 1600 ◦C). X-ray diffraction, scanning electron microscopy and specific surface area measurements were used to analyze the structural and microstructural changes of the as prepared powders heated at different temperatures. Electrical characterization of pure sintered materials was conducted and showed that the incorporation of extra oxide ions, corresponding to the La9.56(SiO4)6O2.34 composition, enhanced the ionic conductivity by one order of magnitude when comparing to that of the La9.33(SiO4)6O2 sample
01 Jan 2008
TL;DR: An environmental burden analyzer for machine tool operations is developed, which can evaluate an NC program from the view point of an environment burden by simulating a cutting process and using emission intensities.
Abstract: We have developed an environmental burden analyzer for machine tool operations, which can evaluate an NC program from the view point of an environment burden by simulating a cutting process and using emission intensities. In this paper, global warming is selected as an impact category and the environmental burden analysis of dry, wet and MQL machining operations are carried out by using this analyzer with consideration of various situations. The influence of the peripheral devices of a machine tool, the spindle and the servo motors, the coolant, the lubricant oil, the cutting tool and the metal chips to global warming is introduced in detail.
01 Dec 2008
TL;DR: A novel class of bit-flipping algorithms for decoding low-density parity-check (LDPC) codes is presented, which are naturally derived from a simple nonlinear objective function.
Abstract: A novel class of bit-flipping (BF) algorithms for decoding low-density parity-check (LDPC) codes is presented. The proposed algorithms, which are called gradient descent bit flipping (GDBF) algorithms, can be regarded as simplified gradient descent algorithms. Based on gradient descent formulation, the proposed algorithms are naturally derived from a simple nonlinear objective function.
Patent•
18 Dec 2008TL;DR: In this paper, a mathematical model of an electric power steering device is used to model the steering feeling of the electric motor and a controller gain adjustment part memorizes a relationship between variation of the characteristic constant and a gain K2 for returning the steering feel to the initial state, and determines a torque instruction value to an electric motor based on the gain K 2.
Abstract: PROBLEM TO BE SOLVED: To provide a control device capable of maintaining steering feeling with sufficient shipping state by automatic adjustment even if an electric power steering device is used for a long period of time. SOLUTION: A parameter presumption unit 120 is obtained by modelling the electric power steering device 1, memorizes a mathematic model including a characteristic constant indicating a physical characteristic of the device, and obtains a steering parameter of the mathematic model from a sensor. Further, the characteristic constant is successively presumed from the obtained value and the mathematic model. A controller gain adjustment part 130 memorizes a relationship between variation of the characteristic constant and a gain K2 for returning the steering feeling to the initial state, determines the gain K2 based on the variation of the characteristic constant and the relationship, and determines a torque instruction value to an electric motor 2 based on the gain K2. Thereby, even if the electric power steering device 1 is used for a long period of time, the steering feeling with sufficient shipping state is maintained by automatic adjustment. COPYRIGHT: (C)2009,JPO&INPIT
TL;DR: The kinetics of the transient heat relaxation of bR after photoexcitation proceeds as a thermal diffusion process, uncorrelated with the photocycle itself, largely improving the quality of the data without affecting the functionality of the bR.
Abstract: We present time-resolved room-temperature infrared difference spectra for the bacteriorhodopsin (bR) photocycle at 8 cm−1 spectral and 5 µs temporal resolution, from 4000 to 800 cm−1. An in situ hydration method allowed for a controlled and stable sample hydration (92% relative humidity), largely improving the quality of the data without affecting the functionality of bR. Experiments in both H216O and H218O were conducted to assign bands to internal water molecules. Room-temperature difference spectra of the L and M intermediates minus the bR ground state (L-BR and M-BR, respectively) were comprehensively compared with their low-temperature counterparts. The room-temperature M-BR spectrum was almost identical to that obtained at 230 K, except for a continuum band. The continuum band contains water vibrations from this spectral comparison between H216O and H218O, and no continuum band at 230 K suggests that the protein/solvent dynamics are insufficient for deprotonation of the water cluster. On the other h...